Exposure to time‐varying electromagnetic fields (EMF) has the capacity to influence biological systems. Our results demonstrate that exposure to time‐varying EMF modeled after the physiological firing frequency of intercellular calcium waves can inhibit proliferation and induce apoptosis in malignant cells. Single exposure of B16‐BL6 cells to a Ca 2+ EMF for 40 min reduced the number of viable cells by 50.3%. Cell imaging with acridine orange and ethidium bromide dye revealed substantial cellular apoptosis, preapoptotic cells, nuclear fragmentation, and large spacing between cells in the Ca 2+ EMF condition when compared to the control condition. The ability of Ca 2+ EMF to influence the proliferation and survival of malignant cells suggests that exposure to specific EMF may function as a potential anticancer therapy.

Honey has become popular as a potential treatment for several ailments, including cancer. Honeys from different parts of the world have been shown to have different anti-proliferative, immune-modulatory, and anti-inflammatory actions. Yemeni Sidr honey (YSH) is world-renowned for its anti-inflammatory activity and has been suggested to have anti-cancer activity, although empirical evidence is lacking. We tested three YSH samples by HPLC to show they contained similar sugars and an overlapping group of phenolic and flavonoid components, as described previously. YSH’s apoptotic and anti-proliferative activities were measured in in vitro models of cancer growth. The treatment of breast cancer cell lines (MDA-MB-231 and MCF-7), a cervical cancer cell line (HeLa), and mouse melanoma cells (B16-BL6) with 1% (w/v) YSH in media for 48–72 h almost completely inhibited cell proliferation and promoted cell apoptosis. In contrast, a non-malignant HBL-100 cell line was more resistant to treatment with YSH. This suggests that YSH may be a good candidate as an anti-cancer treatment, which requires further study.

Differences in the baseline levels of serum cytokines or in single-nucleotide polymorphisms (SNPs) in cytokine genes may be useful to predict outcomes for patients being treated for metastatic breast cancer. We have measured the plasma levels and characterized individual SNPs for IL-1RA, IL-1β, IL-2, IL-6 and TNFα in 130 patients with metastatic breast cancer treated with high-dose chemotherapy. Patients were treated with high-dose cyclophosphamide (Group 1, 74 patients) or high-dose paclitaxel-containing regimens (Group 2, 56 patients). A high plasma level of IL-1RA and a SNP in the IL-1RA gene indicated a better prognosis for patients in Group 1 (but not Group 2). However, the level of plasma IL-1RA did not correlate with the SNP genotype. A high plasma level of IL-6 or TNFα indicated a poorer outcome for patients in Group 1 although the SNP genotypes for the IL-6 and TNFα SNPs were not associated with differences in outcome. The plasma levels of IL-1β and IL-2 and the genotype of the IL-1β SNPs did not indicate differences in outcome. Although, individually, plasma levels of cytokine or “risk” SNP genotypes may not indicate outcome, in combination there was an increased trend to predict outcome for patients treated with high-dose cyclophosphamide but not high-dose paclitaxel. These results suggest that the immune cytokines may be useful as prognostic biomarkers in the treatment of patients with metastatic breast cancer treated with different types of chemotherapy.

Uncaria tomentosa is a medicinal plant native to Peru that has been traditionally used in the treatment of various inflammatory disorders. In this study, the effectiveness of U. tomentosa as an anti-cancer agent was assessed using the growth and survival of B16-BL6 mouse melanoma cells. B16-BL6 cell cultures treated with both ethanol and phosphate-buffered saline (PBS) extracts of U. tomentosa displayed up to 80% lower levels of growth and increased apoptosis compared to vehicle controls. Treatment with ethanolic extracts of Uncaria tomentosa were much more effective than treatment with aqueous extracts. U. tomentosa was also shown to inhibit B16-BL6 cell growth in C57/bl mice in vivo. Mice injected with both the ethanolic and aqueous extracts of U. tomentosa showed a 59 ± 13% decrease in B16-BL6 tumour weight and a 40 ± 9% decrease in tumour size. Histochemical analysis of the B16-BL6 tumours showed a strong reduction in the Ki-67 cell proliferation marker in U. tomentosa-treated mice and a small, but insignificant increase in terminal transferase dUTP nick labelling (TUNEL) staining. Furthermore, U. tomentosa extracts reduced angiogenic markers and reduced the infiltration of T cells into the tumours. Collectively, the results in this study concluded that U. tomentosa has potent anti-cancer activity that significantly inhibited cancer cells in vitro and in vivo.

Previous research has demonstrated that pheochromocytoma (PC12) cells treated with forskolin provides a model for the in vitro examination of neuritogenesis. Exposure to electromagnetic fields (EMFs), especially those which have been designed to mimic biological function, can influence the functions of various biological systems. We aimed to assess whether exposure of PC12 cells treated with forskolin to patterned EMF would produce more plasma membrane extensions (PME) as compared to PC12 cells treated with forskolin alone (i.e., no EMF exposure). In addition, we aimed to determine whether the differences observed between the proportion of PME of PC12 cells treated with forskolin and exposed to EMF were specific to the intensity, pattern, or timing of the applied EMF. Our results showed an overall increase in PME for PC12 cells treated with forskolin and exposed to Burst-firing EMF as compared to PC12 cells receiving forskolin alone. No other patterned EMF investigated were deemed to be effective. Furthermore, intensity and timing of the Burst-firing pattern did not significantly alter the proportion of PME of PC12 cells treated with forskolin and exposed to patterned EMF.

The frequencies of circulating myeloid and T-cell populations were correlated with clinical outcome for 88 patients with metastatic breast cancer treated with high-dose chemotherapy. The ability to predict outcome depended on the chemotherapy regimen. The frequency of monocytes indicated outcome for patients treated with cyclophosphamide-based chemotherapy, while the frequency of T cells indicated outcome for patients treated with paclitaxel-based chemotherapy. Background: The frequency of circulating leukocytes has been shown to be a prognostic factor in patients being treated for different types of cancer. In breast cancer, tumor-infiltrating leukocytes may predict patient outcome, but few studies have investigated such associations for circulating leukocytes. Patients and Methods: Multiparametric flow cytometry was used to examine the immunophenotypes of circulating peripheral blood mononuclear cells for 88 patients with metastatic breast cancer, which was then correlated to breast cancer-specific survival. Patients had been treated either with high-dose cyclophosphamide-containing regimens (group 1, n = 51 patients) or high-dose paclitaxel-containing regimens (group 2, n = 37 patients). Results: The frequency of peripheral blood CD14(+) monocytes indicated prognosis for patients in group 1 (but not group 2), while higher levels of CD11c(+) dendritic cells indicated a better prognosis for patients in group 2 (but not group 1). The frequency of a number of different CD4(+) or CD8(+) T cell subtypes also predicted prognosis for patients in group 2. For example, patients in group 2 with a higher frequency of circulating CD4(+) or CD8(+) naive T cells (CD45RA(+)CD95(-)CD27(+)CD28(+)) showed a poorer prognosis. In contrast, T cells were not associated with prognosis for patients in group 1. Conclusion: Circulating leukocytes can predict clinical outcome for patients with breast cancer. Prediction of clinical outcome in this cohort of metastatic breast cancer patients was specific to the type of chemotherapy, and this finding is likely to apply to other therapies.

Flaxseed oil is widely recognized for its exceptional nutritional value, high concentration of fiber-based lignans and large amounts of ω-fatty acids. It is one of a generic group of functional foods that is often taken by cancer patients as a potential treatment. We have examined the anti-cancer effects of flaxseed oil by studying its direct effects on cancer cell growth in vitro. Treatment of a variety of cancer cell lines with flaxseed oil decreased their growth in a dose-dependent manner while non-malignant cell lines showed small increases in cell growth. Cells treated with a mixture of fatty acids, including α-linolenic acid, docosahexaenoic acid, and eicosapentaenoic acid and lignans including enterodiol and enterolactone was also able to decrease the growth of cancer cells. Treatment of B16-BL6 murine melanoma and MCF-7 breast cancer cells with flaxseed oil induced apoptosis as determined by changes in cell morphology, annexin V staining, DNA fragmentation and/or caspase activation. In addition, treatment with flaxseed oil also disrupted mitochondrial function in B16-BL6 and MCF-7 cells. These results indicate that flaxseed oil can specifically inhibit cancer cell growth and induce apoptosis in some cancer cells and suggests it has further potential in anti-cancer therapy.

Background Cancer patients are increasingly seeking out complementary and alternative medicine (cam) and might be reluctant to disclose its use to their oncology treatment team. Often, cam agents are not well studied, and little is known about their potential interactions with chemotherapy, radiation therapy, or biologic therapies, and their correlations with outcomes. In the present study, we set out to determine the rate of cam use in patients receiving treatment at a Northern Ontario cancer centre. Methods Patients reporting for treatment at the Northeast Cancer Centre (necc) in Sudbury, Ontario, were asked to complete an anonymous questionnaire to assess cam use. Changes in cam use before, compared with after, diagnosis were also assessed. Results Patients in Northern Ontario reported significant cam use both before and after diagnosis. However, as a function of the cam type, cam use was greatly enhanced after cancer diagnosis. For example, the number of patients who reported use of biologic products increased to 51.8% after a cancer diagnosis from 15.6% before a cancer diagnosis. Patients reported much smaller changes in the use of alternative medical systems or spiritual therapy after diagnosis. Vitamin use was reported by 66% of respondents, and the number of different cams used correlated significantly with the reported number of vitamins used. Conclusions Use of cam, particularly biologic products, increased significantly after a cancer diagnosis. Further studies are required to examine the effect of cam use on the efficacy and safety of cancer therapies.

This study evaluates whether three definitions of rural and urban residence predict prostate cancer progression. People were classified as urban or rural using three definitions: rural and small town (RST), Beale's rural-urban continuum codes, and the Rurality Index of Ontario (RIO) 2008 score. This was a chart-based cohort study of males with prostate cancer who underwent external beam radiation therapy (EBRT) in the Regional Cancer Program at Health Sciences North in Sudbury, Ontario from 1996 to 2003. Data indicative of each of the three definitions were used as predictors in Cox regression analysis for the period of 1,000 to 3,000 days after initial diagnosis and as the basis for dichotomous strata in a log rank test. Complete data were acquired from 629 charts. There was no significant association between any of the three definitions of rurality and prostate cancer progression. However, a Beale-based dichotomization led to survival differences using the log rank test. Beale stratification was potentially sensitive to relevant differences in populations that were not represented by the other two definitions. Given the moderate correlations between the different rurality scores, there may be merit to considering multiple rurality scores as they may lead to different cancer progression outcomes in some situations.

Background: Uncaria tomentosa (Willd. ex Schult.) DC (Rubiaceae), known as Cat's Claw or Una de gato, is a traditionally used medicinal plant native to Peru. Some studies have shown that U. tomentosa can act as an antiapoptotic agent and enhance DNA repair in chemotherapy-treated cells although others have shown that U. tomentosa enhanced apoptosis. Objective: To determine if treatment with U. tomentosa can significantly enhance cell death in THP-1 cells exposed to ionizing radiation. Materials and Methods: THP-1 monocyte-like cells were treated with ethanolic extracts of U. tomentosa in the presence or absence of bacterial lipopolysaccharide and then exposed to ionizing radiation. Cell proliferation was assessed by MTT and clonogenic assays and the effects on cell cycle measured by flow cytometry and immunoblotting. Changes in cell signaling were determined by immunoblotting and cytokine ELISA and activation of apoptosis measured by caspase activation and DNA fragmentation analysis. Results: Treatment of THP-1 cells with U. tomentosa had a small effect on cell proliferation. However, when the U. tomentosa-pretreated cells were also subjected to 5-9 Gy ionizing radiation, they showed a significant decrease in cell proliferation and increased cellular apoptosis as measured by DNA fragmentation and caspase activation. Treatment with U. tomentosa also decreased the expression of Cyclin E and Cyclin B, key regulators of normal cell cycle progression, and decreased the phosphorylation of various stress-activated, cell survival proteins including p38, ERK, and SAP/JNK kinase. Conclusions: These results suggest that U. tomentosa could be useful in enhancing cell death following anticancer therapies including ionizing radiation.

Exposure to specific electromagnetic field (EMF) patterns can affect a variety of biological systems. We have shown that exposure to Thomas‐EMF, a low‐intensity, frequency‐modulated (25–6 Hz) EMF pattern, inhibited growth and altered cell signaling in malignant cells. Exposure to Thomas‐EMF for 1 h/day inhibited the growth of malignant cells including B16‐BL6 mouse melanoma cells, MDA‐MB‐231, MDA‐MB‐468, BT‐20, and MCF‐7 human breast cancer and HeLa cervical cancer cells but did not affect non‐malignant cells. The Thomas‐EMF‐dependent changes in cell proliferation were mediated by adenosine 3′,5′‐cyclic monophosphate (cAMP) and extracellular‐signal‐regulated kinase (ERK) signaling pathways. Exposure of malignant cells to Thomas‐EMF transiently changed the level of cellular cAMP and promoted ERK phosphorylation. Pharmacologic inhibitors (SQ22536) and activators (forskolin) of cAMP production both blocked the ability of Thomas‐EMF to inhibit cell proliferation, and an inhibitor of the MAP kinase pathway (PD98059) was able to partially block Thomas‐EMF‐dependent inhibition of cell proliferation. Genetic modulation of protein kinase A (PKA) in B16‐BL6 cells also altered the effect of Thomas‐EMF on cell proliferation. Cells transfected with the constitutively active form of PKA (PKA‐CA), which interfered with ERK phosphorylation, also interfered with the Thomas‐EMF effect on cell proliferation. The non‐malignant cells did not show any EMF‐dependent changes in cAMP levels, ERK phosphorylation, or cell growth. These data indicate that exposure to the specific Thomas‐EMF pattern can inhibit the growth of malignant cells in a manner dependent on contributions from the cAMP and MAP kinase pathways. Bioelectromagnetics. 39;217–230, 2018. © 2017 Wiley Periodicals, Inc.

Tobacco smoking is recognized as a global pandemic resulting in 6 million deaths per year. Despite a variety of anti-smoking products available to aid with tobacco cessation, the majority of people who attempt to quit smoking relapse within 6 months due to the addictive nature of nicotine. An immunotherapy approach could offer a promising treatment option by inducing a potent selective antibody response against nicotine in order to block its distribution to the brain and its addictive effects in the central nervous system. Our nicotine vaccine candidate was administered intranasally using the Neisseria meningitidis serogroup B Adjuvant Finlay Proteoliposome 1 (AFPL1) as a part of the delivery system. This system was designed to generate a robust immune response by stimulating IL-1β production through Toll-like receptor 4 (TLR4), a potent mechanism for mucosal immunity. The vaccine induced high antibody titers in mice sera in addition to inducing mucosal antibodies. The efficacy of our vaccine was demonstrated using in vivo challenge experiments with radioactive [(3)H]-nicotine, followed by an analysis of nicotine distribution in the lung, liver, blood and brain. Our results were encouraging as the nicotine concentration in the brain tissue of mice vaccinated with our candidate vaccine was four times lower than in non-vaccinated controls; suggesting that the anti-nicotine antibodies were able to block nicotine from crossing the blood brain barrier. In summary, we have developed a novel nicotine vaccine for the treatment of tobacco addiction by intranasal administration and also demonstrated that the AFPL1 can be used as a potential adjuvant for this vaccine design.

Exposure to low intensity, low frequency electromagnetic fields (EMF) has effects on several biological systems. Spatiotemporal characteristics of these EMFs are critical. The effect of several complex EMF patterns on the proliferation of B16‐BL6 mouse melanoma cells was tested. Exposure to one of these patterns, the Thomas‐EMF, inhibited cell proliferation and promoted calcium uptake. The Thomas‐EMF is coded from a digital‐to‐analog file comprised of 849 points, which provides power to solenoids and can be set to alter timing, intensity, and duration of variable EMF. Setting the point duration to 3 ms generated a time‐varying EMF pattern which began at 25 Hz and slowed to 6 Hz over a 2.5 s repeat. Exposing B16‐BL6 cells to Thomas‐EMF set to 3 ms for 1 h/day inhibited cell proliferation by 40% after 5 days, while setting the point duration to 1, 2, 4, or 5 ms had no effect on cell proliferation. Similarly, exposing cells to Thomas‐EMF set to 3 ms promoted a three‐fold increase in calcium uptake after 1 h, while the other timings had no effect. Exposure to Thomas‐EMF for as short as 15 min/day slowed cell proliferation, but exposure for 1 h/day was optimal. This corresponded to the effect on calcium uptake where uptake was detected after 15 min exposure and was maximal by 1 h of treatment. Studies show that the specific spatiotemporal character of EMF is critical in mediating their biological activities. Bioelectromagnetics. 38:165–174, 2017. © 2016 Wiley Periodicals, Inc.

         The physical processes by which specific point duration magnetic fields affect aberrant expressions of living matter may involve non-classical mechanisms. The Aharanov-Bohm voltage for a quantum of energy that is convergent with the quotient of the proton’s magnetic moment to its charge multiplied by the viscosity of water at homeostatic temperatures applied across the distance of O-H bonds in conjunction with its phase modulation is about ±4.3 V. Application of frequency shifting, temporally-patterned magnetic fields produced by 3 ms point durations at average intensities of ~28 mG (that are equivalent to Nernst thresholds for plasma membranes) generated through optocoupler light emitting diodes produced the strongest inhibition of malignant cells growth when the pre-coupler value for the circuit maintenance was ±4.3 V compared to increments of voltage below or above this value. Spatial expansion of the effective zone for growth diminishment also occurred with this pre-voltage level. These results indicate that phase modulation of the electrons mediating cellular molecular pathways may be central to the etiology and potential treatment of malignant cells but not  for normal cells’ dynamics. Consideration of quantum effects rather than classical electromagnetic theory may be a more effective strategy for impeding the physical bases for the molecular pathways that define malignant cells.

The physical processes by which specific point duration magnetic fields affect aberrant expressions of living matter may involve non-classical mechanisms. The Aharanov-Bohm voltage for a quantum of energy that is convergent with the quotient of the proton’s magnetic moment to its charge multiplied by the viscosity of water at homeostatic temperatures applied across the distance of O-H bonds in conjunction with its phase modulation is about ±4.3 V. Application of frequency shifting, temporally-patterned magnetic fields produced by 3 ms point durations at average intensities of ~28 mG (that are equivalent to Nernst thresholds for plasma membranes) generated through optocoupler light emitting diodes produced the strongest inhibition of malignant cells growth when the pre-coupler value for the circuit maintenance was ±4.3 V compared to increments of voltage below or above this value. Spatial expansion of the effective zone for growth diminishment also occurred with this pre-voltage level. These results indicate that phase modulation of the electrons mediating cellular molecular pathways may be central to the etiology and potential treatment of malignant cells but not for normal cells’ dynamics. Consideration of quantum effects rather than classical electromagnetic theory may be a more effective strategy for impeding the physical bases for the molecular pathways that define malignant cells.

Increased emissions of photons or shifts in spectral power densities of photons have been reliably measured from malignant cells compared to non-malignant cells. Previous experiments have shown that specific wavelengths within the visible spectrum emitted from melanoma cells were associated with the activation or inhibition of specific molecular structures or pathways. To discern if numbers of photons could differentiate the dynamic state of a critical protein (enzyme), Protein Kinase A (PKA), melanoma cells were transfected with either catalytic subunits, regulatory subunits, or a mutant dominant negative PKA. Compared to typical melanoma cells those transfected with the regulatory subunit exhibited a marked (10 to 100) increase in photon emissions for several hours. The small but significant increase in photon emissions from cells transfected with the catalytic subunit was more brief (first 20 min) and less intense. Photon emissions from cells transfected with inhibitory components did not differ from typical melanoma cells. The vectorial characteristics of the photon emissions were sufficient to clearly differentiate activation of various components of PKA domains.

Previously published findings have documented increased breast cancer risks associated with the nursing profession. The aim of the present study was to assess whether an increased risk of breast cancer was associated with nursing in a population-based case–control breast cancer study of women in Northeastern Ontario, Canada.

This experiment was designed to test the effects of an extremely low frequency electromagnetic field (ELF-EMF) generator (‘Resonator’) compared with strong static magnetic fields on cellular viability, and to illustrate the importance of dynamic patterns of fields compared with static patterns.

Spring water but not double-distilled water was exposed, in darkness, to a temporally patterned weak magnetic field that has been shown to affect planarian behavior and slow the rate of cancer cell proliferation. Exposure to the magnetic field caused a reliable shift in the peak (longer) wave-length of ~10 nm for fluorescence emissions and a ~20% increase (~100 counts) in fluorescence intensity. Spectral analyses verified a shift of 5 and 10 nm, equivalent to ~1.5 × 10-20 J “periodicity” across the measured wavelengths, which could reflect a change in the an intrinsic energy as predicted by Del Giudice and Preparata and could correspond to two lengths of O-H bonds. Wrapping the water sample containers during exposure with copper foil, aluminum foil, or plastic altered these fluorescent profiles. The most conspicuous effect was the elimination of a ~280 nm peak in the UV-VIS emission spectra only for samples wrapped with copper foil but not aluminum or plastic. These results suggest that weak magnetic fields produce alterations in the water-ionic complexes sufficient to be reliably measured by spectrophotometry. Because the effect was most pronounced when the spring water was exposed in darkness and was not disturbed the role of thixotropic phenomena and Del Giudice entrapment of magnetic fields within coherent domains of Pollack virtual exclusion zones (EZ) may have set the conditions for subsequent release of the energy as photons.